Background: Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), Porcine Circovirus Type 2 (PCV2) and Hepatitis E virus (HEV) are common and economically important viral disease causative agents detected in pig oral fluid (OF), faeces and serum at some infection stages. The purpose of this study was to detect PRRSV, PCV2 and HEV on six pig farms to determine which of the three sample types, OF, faeces or serum is appropriate for the diagnosis of these viruses in different pig categories.The following pig categories were included: 5 weeks-old (w/o), 7 w/o, 9 w/o, 11 w/o weaners, fatteners and breeding sows. Pursuant to the preliminary detection of each pathogen at the selected farms, OF samples, faeces, serum pools and 10 individual sera were examined, using PCR, for each age category. If any of the viruses were found in pools of faeces and OF, then faeces and OF from positive farms were tested separately for each pig category. The viral nucleic acids were detected using RT-PCR, PCR and real-time RT-PCR, for PRRSV, PCV2 and HEV respectively.Results: PRRSV and HEV were detected on one farm and PCV2 on three others, positive results being more often obtained from the OF than from the faeces of the same animals. Ten individual serum samples from pigs from the same group of animals were also tested. The viruses were detected in almost all individual sera and OF in the same pig category with some exceptions: PRRSV was detected in the OF of fatteners but was absent in their sera; on Farm 2, PCV2 was detected in sera of 11 w/o pigs and fatteners but absent in group samples of their OF and, vice versa, in case of 9 w/o animals; HEV was detected in the OF of the youngest, 5 w/o weaners and absent in sera of the same age group.Conclusions: The primary finding of the study is that OF is a welfare-friendly, non-invasive and highly efficient matrix for pathogen detection, thus evidencing the usefulness of pig OF as a matrix in which each of the three viruses considered can be detected with the highest probability.

African Swine Fever (ASF) is persistently spreading and hindering pork production in Europe. Slovenia is one of the last countries in Central Europe without a confirmed ASF case in domestic pigs or in wild boar. The aim of this study was to assess the current biosecurity implementation on different types of pig farms. Internal and external biosecurity status was determined in 17 commercial (CF), 15 non-commercial (NC), and 15 outdoor (O) farms. Data were collected using the Biocheck.UGent questionnaire and assessed in combination with the latest information on the wild boar population in Slovenia. Biosecurity was compared between farm types based on the assessment of 12 subcategories. Statistically significant differences (p < 0.05) were found in six subcategories: (i) purchase of pigs and semen, (ii) visitors and farmworkers, (iii) vermin and bird control, (iv) finishing unit, (v) measures between compartments and use of equipment, and (vi) cleaning and disinfection. The highest total biosecurity score (0–100%) was determined on CF with 64.59 ± 16.47%, followed by NC with 55.73 ± 10.67%, and O with 48.47 ± 8.20%. The density of the wild boar population was estimated from the number of wild boars per km² per year, with 3 or more hunted wild boars per unit representing the highest density. Geolocation of farms on the wild boar population map showed that two O farms are at high risk and seven farms (1 O, 5 NC, and 1 CF) are at medium risk for disease transmission from wild to domestic pigs. Biosecurity measures must be tightened in some subcategories, especially in areas with a high density of wild boar.

Animal welfare is important; therefore, veterinarians and other animal welfare experts try to use different tools for pig welfare assessment. Several welfare protocols are available for pig welfare assessment, and one of the most used is Welfare Quality (WQ) protocol^®. Elevated values of cortisol can be indicative of stress and, therefore, poor welfare. Our aim was to assess the correlation between serum cortisol levels from individual samples and oral fluid cortisol levels in group samples with the grades received for pig welfare using the WQ protocol^®. Samples were taken at six different commercial pig farms. Animals were divided into age-dependent categories: 5 weeks old (w/o); 7 w/o; 9 w/o; 11 w/o weaners; fatteners; and breeding sows (10 pigs/category). Cortisol was determined in individual sera and group samples of oral fluid (OF), and was compared to values considered to be physiological. Based on WQ protocol^® answers, five farms’ welfare level was deemed acceptable, and one was enhanced. Four out of 29 sera and 5 out of 30 OF samples were considered physiological, while in most other samples it was elevated. The correlation between cortisol levels in sera, OF, and WQ protocol^® scores was not statistically significant. The cortisol level in OF should be just one of the welfare indicators, i.e., alongside the WQ protocol^® filled out by a welfare expert.

The first case of ASF in a domestic pig population in Serbia was confirmed in 2019 in a backyard population. Today, outbreaks in wild boar and, more importantly, in domestic pigs are still occurring, although the government measures for ASF prevention are in place. The aim of this study was to determine critical risk factors and identify the possible reasons for ASF introduction into different extensive pig farms. The study was conducted on 26 extensive pig farms with confirmed ASF outbreaks, with data collected from beginning of 2020 to the end of 2022. Collected epidemiological data were divided into 21 main categories. After identifying specific values of variables as critical for ASF transmission, we identified nine important ASF transmission indicators as those variables for which at least 2/3 of the observed farms reported values critical for ASF transmission. Among them were type of holding, distance to hunting ground, farm/yard fencing, and home slaughtering; however, the hunting activity of pig holders, swill feeding, and feeding with mowed green mass were not included. We represented the data in the form of contingency tables to study associations between pairs of variables using Fisher’s exact test. All pairs of variables in the group including type of holding, farm/yard fencing, domestic pig–wild boar contact, and hunting activity were significantly related; hunting activity of pig holders, holding pigs in backyards, unfenced yards, and domestic pig–wild boar contact were observed on the same farms. Free-range pig farming led to observed domestic pig–wild boar contact on all farms. The identified critical risk factors need to be strictly addressed to prevent the further spread of ASF to extensive farms and backyards in Serbia and elsewhere.

Background: Porcine circovirus type 3 is the most recently discovered porcine circovirus, and an emerging pathogen. In this study the status of its presence on some Slovenian farms is reported. The effectiveness of the vaccine against porcine circovirus type 2 was assessed against porcine circovirus type 3. Group samples of oral fluid, faeces and individual serum samples were taken from six different pig categories and tested for presence of viral DNA, using both real time and conventional PCR. Positive samples were subjected to direct Sanger sequencing. Nucleotide sequences were analyzed and compared to GenBank PCV3 sequences. Results: Positive samples were sent for genome sequencing, which confirmed the presence of virus in all different pig categories on five farms. A high to moderate correlation of strong statistical significance was found between individual serum samples, oral fluid and faeces. Slovenian PCV3 was found to be distributed in a way similar to that of other countries. Slovenian PCV3 nt sequences are highly related, sharing more than 99.5% nt identity. On one farm a commercially available vaccine against porcine circovirus type 2 was used on 3-week-old pigs. It did not affect the presence of porcine circovirus type 3 in oral fluid or sera of any of the seven age groups of pigs, each with two control groups. Conclusions: The results constitute the first discovery of the virus in Slovenia. Genome sequencing has revealed a high degree of similarity between Slovenian and GenBank isolates.

Summary: African swine fever (ASF) is a serious disease of domestic and feral swine mainly because of grave clinical course of the disease and its strong socio-economic impact. Not only there is an extremely high mortality (95–100%) around the time of the first outbreak, live-animal, pork meat and other pork product trade is strongly affected on regional and international level. ASF is a species-specific disease; strength of the clinical signs depends on virulence of a viral strain. Exacerbation of body temperature and dysfunction of respiratory system and gastrointestinal tract are most common occurrences. Lesions can be mostly classed as congestive or hemorrhagic. Epidemiologically, ASF varies substantially among regions, countries and continents due to wide panel of different virus genotypes, population of feral swine and other reservoirs of disease, geographical characteristics and social habits of people. ASF is endemic in Sub-Saharan Africa where soft ticks (Argaside) and chronically infected feral swine population serve as reservoirs. It also became endemic in most parts of Russian Federation. Since 1978, ASF is endemic on Sardinia isle, as it was eradicated in all other parts of Europe. First outbreak in Georgia in 2007 has apparently served for transmission to other eastern European countries. Slovenia is currently still ASF-free but disease has already reached some relatively nearby countries (Poland, Romania, Moldova, Czech Republic and Hungary). There is no anti-ASF vaccine currently available ad specific treatment has not been described. Therefore rapid diagnostics and implementation of strict biosecurity measures play a key role in prevention of further transmission.